Aquilaria agallocha can produce fragrant agarwood used for incense, traditional medicine and other products. An efficient plant regeneration system was established via organogenesis from shoots developed from seedlings of Aquilaria agallocha. Shoots generated many buds on MS medium supplemented with 1.3 micromol/L BA (6-benzylaminopurine) in the first 7 weeks, and the buds elongated on MS medium with 1.3 micromol/L BA+0.5 micromol/L NAA (naphthaleneacetic acid) in another 7 weeks, 2.3 shoots 2 cm in length per explant were obtained within 14 weeks. Plantlets were rooted on 1/2 MS medium after being immersed in 5 micromol/L NAA for 48 h, 96.7% of the roots grew up two weeks later. All plantlets that survived acclimatization grew well in the pots.
Agriculture ; methods ; Cell Culture Techniques ; methods ; Forestry ; methods ; Phytotherapy ; Plant Growth Regulators ; metabolism ; Plant Shoots ; drug effects ; growth & development ; Survival Analysis ; Survival Rate ; Thymelaeaceae ; drug effects ; growth & development

Objective To explore the value of 11C-choline PET/CT in patients with hepatic spaceoccupying lesions that have an indeterminate diagnosis by 18F-fluorodeoxyglucose (FDG) PET/CT. Methods A total of 25 liver masses in 20 patients with an indeterminate diagnosis based on 18F-FDG PET/CT were enrolled. Regional 11C-choline PET/CT scan was performed in all of the patients. Lesions with intense 11C-choline uptake were considered as positive. The semiquantitative maximum standardized uptake value(SUVmax) was measured and the tumor-to-liver (T/L) radioactivity ratio was calculated. The Mann-Whitney test,Kruskal-Wallis test and crosstabs x2-test were performed by using SPSS version 11.5. Results Of the 25 lesions,21 were proven to be hepatocellular carcinomas (HCC),3 hemangiomas,and 1 parasitic granuloma. The sensitivity of 11C-choline PET/CT for the detection of HCC was 66.7% (14/21). 11C-choline PET/CT had a higher sensitivity for well differentiated HCC than moderately and poorly differentiated HCC on a patient basis (8/9 vs 2/5,respectively). There were significant differences of 11C-choline T/L ratios between the HCC positive group,HCC negative group and benign lesion group ( 1.70 ± 0.35,0.86 ± 0.15,and 0.36 ± 0.18,x2 = 19.00,P ＜0.01 ). The lesion size and alphafetoprotein (AFP) level between the HCC positive and negative groups had no significant difference respectively ( Mann Whitney U = 39.00,P ＞0.05,and U=16.00,P＞0.05,respectively). Conclusions 11C-choline is complementary to 18F-FDG PET/CT for the detection of HCC,especially for well differentiated HCC.

As products of interaction of time and space, geoherbs, which are essential parts of Chinese Materia Medica, were characterized in different morphology, unique habitat, continuous and changeable sites. The main fields in molecular mechanism of geoherbs focus on: biodiversity and molecular identification, genetic different and evolutionary genetics, geo-variation and environmental adaptation, germplasm and aimed genus choosing, expression and control of functional gene, gene transfer and bio-safety evaluation. The main tasks are to discover the genetic variation at molecular level, ascertain the molecular characteristics of geoherbs and the effect of environment on gene expression of geoherbs, confirm the genetic factors attribute to the forming of geoherbs, and find out the genetic basis of geoherbs at individual level and population level, respectively. This paper pointed out that the essential of geoherbs is continuers quantities variation at population level, geoherb's populations are different in gene frequency with the others'; geohersm are quantitative trait loci (QTL) controlled by multi - gene or combination with multiple-gene and major gene at individual level. It is very important to pay more attention to the scale effect of geoherbs, refer the theories and methods of quantities genetic, and concern more about the interaction of environment and gene in geoherbs' molecular mechanism research.
Drugs, Chinese Herbal ; adverse effects ; metabolism ; pharmacology ; Geography ; Plants, Medicinal ; classification ; genetics ; growth & development ; Quantitative Trait Loci

Based on the outpatient interview and literature review,the initial framework of the outpatient experience of human caring scale was formed with 9 dimensions of outpatient process.The research aim was to improve the scale by Delphi method.Sixteen experts in medical management,human caring or medical education were invited to evaluate the importance of the dimensions and items of the scale and provided some expertise via filling out the Delphi consultation questionnaires twice in the consulting round.In the first round,the recovery rate showing the experts' positivity was 80％;the coefficient of reliability (Cr) ascertaining the authority of the evaluation was 0.92;the mean and full mark ratios responding the concentration of the evaluation were 2.88-4.94 and 6.25％-93.75％ respectively;the coefficients of variation (CV) and the Kendall's W determining the concordance of the evaluation were 5.06％-52.15％ and 0.21-0.24 respectively.In the second round,the recovery rate was 93.75％;the Cr was 0.93;the mean was 3.93-4.93;the full mark ratios were 26.67％-93.33％;the Kendall's W was 0.14-0.31,the CV was 5.25％-23.61％.Via the two-round Delphi study,the scale that included 10 dimensions and 61 items has been improved.Ten dimensions are pre-hospital medical service,guidance,registration,waiting,diagnosis & treatment,paying,inspection & assay,medicine receiving,therapy/injection/transfusion and global evaluation.It was concluded that Chinese scholars have paid high attention to human caring and outpatient experience.The experts have given high agreements about the dimensions which were established with Chinese outpatient process.The dimensions are different from the similar researches about outpatient experience study.In the future,it is necessary to survey the outpatients to test the construct validity,internal consistency reliability and others of the scale to improve the scale.

Objective To investigate the pharmacokinetics of cyclosporine(CsA) in type 1 diabetes of rats.Methods Rats with type 1 diabetes were induced by intraperitoneal administration of 65 mg· kg-1 streptozotocin (STZ).Pharmacokinetics of CsA (10 mg· kg-1) was compared between diabetic and normal rats on the 5 weeks after STZ injection.Concentration of CsA in whole blood was measured by enzyme multiplied immunoassay technique.Results Type 1 diabetes rats were successfully induced as the fasting blood glucose levels exceeded 11.1 mmol · L-1 after STZ injection for 1 week.Beside glucose,total cholesterol (TC) and triacylglycerol (TG) in serum of diabetic rats were significantly higher than those in normal rats after STZ injection for 5 weeks.Unlike normal rats,the pharmacokinetics of CsA in diabetic rats exhibited a double peak after oral administration.The Tmax and Cmax were lower in diabetic rats than normal rats,although without statistical differences as follows:Tmaxwas (3.3±1.6),(3.2 ±2.5) h,Cmax was (579.0±208.5),(453.0 ± 104.8) ng· mL-1 in normal and diabetic rats.Compared with normal rats,AUC and t1/2 of CsA in diabetic rats decreased significantly,which were 0.51 and 0.70 times those of normal rats,respectively;AUC was (7343.2 ± 2333.7),(3729.7 ± 1106.6) h ·ng · mL-1,t1/2 was (8.5 ± 1.5),(6.0 ± 0.9) h in normal and diabetic rats.Conclusion The oral pharmacokinetics of CsA was significantly different in type 1 diabetic rat model compared to normal controls,which should be taken into consideration in clinical medication.

OBJECTIVETo explore a method which can remove the gastric mucus in order to prepare mucous membrane single cell suspension for the research of cytomics.

METHODSEnzymology was used to remove the mucus gel and to separate mucous layer from the normal fresh gastric tissue. The mucous layer was broken to prepare single cell suspension with machine method. Expression of major cyclins in mucous layer cells was examined by cytoimmunochemistry, flow cytometry(FCM) and confocal microscopy.

RESULTSThe 0.1% pepsin could dissolve the mucus gel and 1.2-2.4 U/L dispase could separate the mucous layer completely. The single mucous cell suspension was prepared successfully. FCM results from mucous single cell suspension revealed that expression of cyclin D(3), B(1) was obvious, that of cyclin D(2) was weak and that of cyclin D(1), A, E was the least. Similar results were found with confocal microscopy.

CONCLUSIONSSingle cell suspension from mucous layer can be easily prepared by pepsin and dispase. Cyclins schedule expression in vivo is different from cyclins schedule expression in vitro.

Cell Line ; Cell Proliferation ; Cyclins ; metabolism ; Flow Cytometry ; Gastric Mucins ; metabolism ; Gastric Mucosa ; cytology ; metabolism ; Humans ; Mucous Membrane ; cytology ; metabolism

OBJECTIVETo investigate whether low molecular weight heparin (LMWH) may suppress the expression and secretion of vascular endothelial growth factor (VEGF) from tumor cells in vitro and inhibit the VEGF-induced proliferation of human tumor vascular endothelial cells.

METHODSHuman lung cancer cell line A549, human liver cancer cell line HepG2, human colon carcinoma cell lines HCT116 and HCT8 were used in this study. The expression levels of VEGF and TNF-alpha (tumor necrosis factor-alpha) in the tumor cells with or without pretreatment of LMWH/heparin were measured by standard sandwich ELISA technique. The VEGF mRNA level of HepG2 cells cultured with or without LMWH/heparin was determined by RT-PCR and real time PCR. Human umbilical vein endothelial cells (HUVEC) were cultured in tissue culture medium (TCM) with or without LMWH/heparin for 3 days. Then non-radioactive cell proliferation assay (MTS) kit and cell cycle assay by flow cytometry were performed to measure the proliferation of HUVEC.

RESULTSThe VEGF levels in the control, LMWH, and heparin groups of the pulmonary adenocarcinoma cell line A549 were (1045.89 +/- 165.30) pg/ml, (782.45 +/- 67.17) pg/ml and (916.54 +/- 71.25) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups of the colon adenocarcinoma cell line HCT116 were (955.76 +/- 51.14) pg/ml, (822.89 +/- 142.39) pg/ml and (951.77 +/- 188.22) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the colon adenocarcinoma cell line HCT8 were (1290.62 +/- 41.23) pg/ml, (1063.34 +/- 63.82) pg/ml and (1257.14 +/- 11.40) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the liver cancer cell line HepG2 were (1083.00 +/- 134.35) pg/ml, (758.00 +/- 84.85) pg/ml and (874.00 +/- 22.62) pg/ml, respectively. The VEGF expression levels in the above mentioned cell lines cultured in TCM were significantly reduced in the LMWH-treated groups compared with that of the control group (P < 0.05). But the level of TNF-alpha in TCM-cultured cells was unaffected by LMWH. The VEGF mRNA was reduced in the LMWH-treated HepG2 cell line. Moreover, TCM exhibited stimulating effect on proliferation of HUVEC and the effect was significantly impaired by LMWH treatment. Flow cytometric analysis revealed that LMWH treatment arrested HUVECs at the G1 phase of cell cycle.

CONCLUSIONLMWH can suppress the expression and secretion of VEGF by tumor cell lines and therefore have a potential inhibiting effect on angiogenesis induced by VEGF.

Adenocarcinoma ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; Endothelial Cells ; cytology ; HCT116 Cells ; Hep G2 Cells ; Heparin ; pharmacology ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; secretion

BACKGROUNDRecent studies have shown that interleukin-3 receptor alpha (CD123) is highly expressed on leukemia stem cells of patients with acute myeloid leukemia, and is correlated with tumor load and poor prognosis. The expression of CD123 may also be high in patients with myelodysplastic syndrome (MDS). In this study, the expression and clinical significance of CD123 and granulocyte colony stimulating factor (G-CSF) receptor (CD114) on the bone marrow cells of patients with MDS were investigated to explore the molecular marker of the malignant clone of MDS.

METHODSForty-two patients with MDS, who were diagnosed in the Hematological Department of General Hospital of Tianjin Medical University from 2008 to 2009, and twelve normal controls were enrolled in this study. Fluorescence activiated cell sorter (FACS) was used to measure the expression of CD123 on CD34(+)CD38(-) cells and CD114 on CD34(+) cells of the bone marrow of these patients and controls and the clinical significance was analyzed. The expression of CD114 on CD123(+)CD34(+)CD38(-) cells was further measured to explore the molecular marker of the malignant clone in MDS.

RESULTSMDS patients displayed significantly higher proportion of CD34(+)CD38(-)/CD34(+) ((14.03 +/- 5.27)%) than normal controls ((7.70 +/- 4.36)%, P < 0.05). The expression rate of CD123(+)CD34(+)CD38(-)/CD34(+)CD38(-) was significantly higher in MDS patients ((48.39 +/- 28.15)%) than that in normal controls ((8.75 +/- 11.71)%, P < 0.01). The expression level of CD123 was significantly correlated with the proportion of bone marrow blasts (r = 0.457, P < 0.05). The expression rate of CD114(+)CD34(+)/CD34(+) was lower in MDS patients ((33.05 +/- 21.71)%) than that in normal controls ((38.99 +/- 19.07)%) but was not statistically significant (P > 0.05). The expression of CD114 on CD123(+)CD34(+)CD38(-) cells ((34.82 +/- 29.58)%) was significantly lower than that on CD123(-)CD34(+)CD38(-) cells ((53.48 +/- 27.41)%) of MDS patients (P < 0.05).

CONCLUSIONSMDS patients displayed higher proportion of CD34(+)CD38(-)/CD34(+) than normal controls. CD123 was highly expressed in the bone marrow of the patients with MDS, significantly correlated with the proportion of bone marrow blasts, and thus might be the marker of MDS malignant clone. CD123(+)CD34(+)CD38(-) cells exhibited lower expression of G-CSF receptors, which might partly explain why MDS clone responds worse to G-CSF in vitro and in vivo.

ADP-ribosyl Cyclase 1 ; metabolism ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; metabolism ; Cells, Cultured ; Female ; Humans ; Interleukin-3 Receptor alpha Subunit ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; metabolism ; Young Adult

OBJECTIVETo explore the value of trisomy 22 ( +22) in the diagnosis of inv(16) acute myeloid leukemia (AML).

METHODSInterphase fluorescence in situ hybridization (FISH) was performed in 18 AML patients with +22. The probe was two-color break apart probe for CBFbeta with SpectrumRed on the centromeric side and SpectrumGreen on the telomeric side. The FISH results were compared with that of R-banding conventional cytogenetics (CC). Multiplex FISH (M-FISH) was used to analyze the relationship of +22 and inv(16).

RESULTSCC revealed inv(16) in none of the 18 AML, with +22, but FISH revealed inv (16) in 11 of them and del( 16) (q22) in one. As CC results, 9 of the 11 cases were sole +22, one complicated with trisomy 8, and one del(16) (q22). Four patients with +22 and inv(16) were analyzed by M-FISH and revealed +22 only.

CONCLUSION+22 can be regarded as an important marker for the diagnosis of inv(16) AML.

Adolescent ; Adult ; Aged ; Chromosome Inversion ; Chromosomes, Human, Pair 16 ; genetics ; Chromosomes, Human, Pair 22 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Trisomy

OBJECTIVETo identify the mutational genotype of three Chinese families with X-linked adrenoleukodystrophy (X-ALD: MIM#300100).

METHODSTotal RNA was extracted from the peripheral blood leukocytes of patients 1, 2 and the mother of patient 3, using RNA blood Mini kit (QIAGEN). After reverse transcription, cDNA was amplified in four overlapping segments. The PCR products were purified and directly sequenced. To confirm the mutations, the genomic DNA was isolated from the patients and their family members using DNA blood isolation kit (MO-BIO) and analyzed by PCR-restrictive digestion or amplification refractory mutation system.

RESULTSThree distinct mutations were detected in the ABCD1 gene of the three pedigrees. A mutation of CCC-->CGC was detected at codon 534 of the ABCD1 gene from patient 1, resulting in the arginine for proline substitution. A change of GGG-->AGG was found at codon 266 of the second patient's gene, accompanied with the replacement of glycine by arginine. A mutation of CGC-->GGC was found at codon 617 in one ABCD1 allele of the third patient's mother, leading to the glycine for arginine substitution. The three mutations were confirmed through restriction analysis or amplification refractory mutation system.

CONCLUSIONThree ABCD1 gene missense mutations were detected in three unrelated Chinese families with X-linked adrenoleukodystrophy, one of which, the mutation (P534R), is novel in Chinese with ALD, and the other two G266R and R617G mutations, have been reported outside China.

ATP Binding Cassette Transporter, Sub-Family D, Member 1 ; ATP-Binding Cassette Transporters ; genetics ; Adrenoleukodystrophy ; genetics ; Child ; Child, Preschool ; Humans ; Male ; Mutation ; Pedigree